ELISA Kit for Vimentin (VIM) Bos taurus; Bovine (Cattle) Sandwich ELISA

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  • ELISA Kit for Vimentin (VIM) Packages (Simulation)
  • ELISA Kit for Vimentin (VIM) Packages (Simulation)
  • ELISA Kit for Vimentin (VIM) Results demonstration
  • SEB040Bo.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Vimentin (VIM) and the recovery rates were calculated by comparing the measured value to the expected amount of Vimentin (VIM) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 79-99 86
EDTA plasma(n=5) 88-102 94
heparin plasma(n=5) 81-104 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Vimentin (VIM) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Vimentin (VIM) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Vimentin (VIM) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 88-99% 80-104% 78-89% 98-105%
EDTA plasma(n=5) 80-105% 92-101% 91-98% 79-97%
heparin plasma(n=5) 96-104% 82-101% 85-99% 83-98%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Vimentin (VIM)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Vimentin (VIM). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Vimentin (VIM). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Vimentin (VIM), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Vimentin (VIM) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Aberrant Vimentin Methylation Is Characteristic of Breast Cancerpubmed:22315367
  • Glyoxalase 1-knockdown in human aortic endothelial cells - effect on the proteome andendothelial function estimates.pubmed:27898103
  • HtrA3 is a cellular partner of cytoskeleton proteins and TCP1α chaperoninPubmed:29477555
  • Growth hormone induces Notch1 signaling in podocytes and contributes to proteinuria in diabetic nephropathyPubmed: 31511328
  • Extracellular vesicles derived from injured vascular tissue promote the formation of tertiary lymphoid structures in vascular allograftsPubmed: 31729155
  • Is Vimentin the Cause or Effect of Obstructive Sleep Apnea Development?Pubmed: 32088750
  • ZNF471 modulates EMT and functions as methylation regulated tumor suppressor with diagnostic and prognostic significance in cervical cancer33566221
  • Metastatic suppression by DOC2B is mediated by inhibition of epithelial-mesenchymal transition and induction of senescence33758996
  • Comparison of Vimentin Levels Between Preeclamptic and Normotensive Pregnant Women
  • Advanced glycation end-products associate with podocytopathy in type II diabetic patients
  • Non-POU Domain-Containing Octamer-Binding (NONO) Protein Stability Regulated by PIN1 is Crucial for Breast Cancer Tumorigenicity Via the MAPK/β …

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